Endogenous bone marrow-derived cells express retinal pigment epithelium cell markers and migrate to focal areas of RPE damage
Endogenous bone marrow-derived cells express retinal pigment epithelium cell markers and migrate to focal areas of RPE damage
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DOI：
10.1167/iovs.06-1015
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发表时间：
2007-09-01
影响因子：
4.4
通讯作者：
Enzmann, Volker
中科院分区：
文献类型：
--
作者：
Li, Yang;Atmaca-Sonmez, Pelin;Enzmann, Volker
目的.本研究旨在探讨骨髓源性细胞（bone marrow derived cells，BMCs）能否在体外被诱导表达视网膜色素上皮（retinal pigment epithelial，RPE）细胞标志物，动员后能否在体内归巢至RPE损伤部位并表达RPE细胞系标志物。将成人RPE细胞与绿色荧光蛋白（GFP）标记的干细胞抗原-1阳性（Sca 1（+））BMCs共培养1、2和3周。研究了细胞形态和RPE特异性标志物和其他视网膜细胞类型标志物的表达。使用碘酸钠（NaIO3）诱导的RPE变性的动物模型，通过粒细胞集落刺激因子、flt3配体或两者将BMC动员到外周循环中。免疫细胞化学用于鉴定和表征C57 BL/6野生型（wt）小鼠和GFP嵌合小鼠视网膜下腔中的BMC。在体外，BMCs从圆形变成扁平的多边形细胞，并表达细胞角蛋白，RPE 65，和小眼转录因子（MITF）时，在直接细胞-细胞接触与RPE共培养。在体内，在视网膜下腔中的BMC被鉴定为Sca-1（+）或c-kit（+）细胞。它们还被GFP和RPE 65或MITF双重标记。这些细胞在RPE损伤的焦点区域的Bruch膜上形成单层。因此，似乎当动员到外周循环中时，BMC可以归巢到RPE损伤的焦点区域并表达RPE谱系的细胞标志物。使用内源性BMCs替代受损的视网膜组织为眼科的细胞替代疗法开辟了新的可能性。
PURPOSE. The aim of the present study was to investigate whether bone marrow-derived cells (BMCs) can be induced to express retinal pigment epithelial (RPE) cell markers in vitro and can home to the site of RPE damage after mobilization and express markers of RPE lineage in vivo.METHODS. Adult RPE cells were cocultured with green fluorescence protein (GFP)-labeled stem cell antigen-1 positive (Sca1(+)) BMCs for 1, 2, and 3 weeks. Cell morphology and expression of RPE-specific markers and markers for other retinal cell types were studied. Using an animal model of sodium iodate (NaIO3)-induced RPE degeneration, BMCs were mobilized into the peripheral circulation by granulocyte-colony stimulating factor, flt3 ligand, or both. Immunocytochemistry was used to identify and characterize BMCs in the subretinal space in C57BL/6 wild-type (wt) mice and GFP chimeric mice.RESULTS. In vitro, BMCs changed from round to flattened, polygonal cells and expressed cytokeratin, RPE65, and microphthalmia transcription factor ( MITF) when cocultured in direct cell-cell contact with RPE. In vivo, BMCs were identified in the subretinal space as Sca-1(+) or c-kit(+) cells. They were also double labeled for GFP and RPE65 or MITF. These cells formed a monolayer on the Bruch membrane in focal areas of RPE damage.CONCLUSIONS. Thus, it appears that BMCs, when mobilized into the peripheral circulation, can home to focal areas of RPE damage and express cell markers of RPE lineage. The use of endogenous BMCs to replace damaged retinal tissue opens new possibilities for cell replacement therapy in ophthalmology.